LPS로 자극된 RAW264.7 대식세포에 대한 뚝지알에서 추출한 중성지질, 당지질, 그리고 인지질의 항염증 효과
Anti-inflammatory Properties of Neutral lipids, Glycolipids, and Phospholipids isolated from Aptocyclus ventricosus Eggs in LPS-stimulated RAW264.7 Cells
- 주제(키워드) 도움말 Anti-inflammatory , Aptocylus ventricosus eggs , RAW264.7 Cells
- 발행기관 강릉원주대학교 일반대학원
- 지도교수 도움말 박우정
- 발행년도 2024
- 학위수여년월 2024. 2
- 학위명 석사
- 학과 및 전공 도움말 일반대학원 웰니스바이오산업학과
- 세부분야 해당없음
- 실제URI http://www.dcollection.net/handler/kangnung/000000011769
- UCI I804:42001-000000011769
- 본문언어 영어
초록/요약 도움말
Aptocyclus ventricosus (Smooth lumpsucker) eggs were harvested for the extraction of total lipids, which were subsequently separated into neutral lipids, glycolipids, and phospholipids. The current study evaluated the fatty acid profiles of neutral lipids, glycolipids, and phospholipids from A. ventricosus eggs, as well as their anti-inflammatory properties via cytotoxicity, NO production, the mRNA expression of immune-associated genes, protein expression of immune signaling pathways, and the expression of cell surface molecules against RAW264.7 macrophages. Firstly, the compositions of fatty acids of three lipids were identified by gas chromatography (GC) with flame-ionization detection (FID) analysis. The neutral lipids were found the highest monosaturated fatty acids (MUFAs) at 46.08%, while the glycolipids, and phospholipids were found the highest polyunsaturated fatty acids (PUFAs) at 41.61% and 39.31%, respectively. The main fatty acids in the three lipids were palmitic acid (17.04–26.91%), oleic acid (12.17–24.60%), eicosapentaenoic acid (EPA 7.76–19.28%), and IX docosahexaenoic acid (DHA 8.33–17.98%). Secondly, the anti-inflammatory effects of fractionated lipids isolated from A. ventricocus eggs were determined in LPS-stimulated RAW264.7 cells. Cells were pre-treated with each lipid at various concentrations (100, 200, 300, and 400 μg/mL) and stimulated by LPS (1 μg/mL). According to the results, three lipids dramatically decreased the amount of NO produced in response to LPS stimulation and the expression of immune-related genes such as iNOS, COX-2, IL-1β, IL-6, TNF-α, and IL-4. The phosphorylated NF-κB-p65 and phosphorylated MAPK proteins (p38, JNK, and ERK 1/2) were down-regulated by A. ventricocus lipids, suggesting the three lipids stimulated by LPS exert anti-inflammatory activity via preventing NF-κB and MAPK activation. In addition, LPS-stimulated CD40 expression was reduced by A. ventricocus fractionized lipids in RAW264.7 cells depending on the concentration of the lipids. Therefore, these results indicate that the neutral lipids, glycolipids, and phospholipids isolated from A. ventricocus eggs could be considered as a potential lipid source for inflammatory diseases.
more목차 도움말
Table of contents I
List of Tables IV
List of Figures V
Acknowledgements VII
Abstract VIII
Chapter 1. Literature Review 1
1. Immune response 1
2. Lipids 3
3. Aptocyclus ventricosus 6
4. References 8
Chapter 2. Fatty acid compositions of neutral lipids, glycolipids, and phospholipids from Aptocyclus ventricosus eggs 14
Abstract 14
1. Introduction 15
2. Materials and methods 16
2.1 Preparation of Aptocyclus ventricosus eggs 16
2.2 Lipid extraction of A. ventricosus eggs 16
2.3 Lipid separation of total lipids from A. ventricosus eggs 16
2.4 Preparation for fatty acid methyl esters (FAMEs) from neutral lipids, glycolipids, and phospholipids 17
2.5 Analysis of the fatty acid compositions 17
2.6 Statistical analysis 18
3. Results and discussion 18
4. Conclusions 23
5. References 23
Chapter 3. Neutral lipids, glycolipids, and phospholipids from A. ventricosus eggs attenuate cellular inflammation in LPS-stimulated macrophages 27
Abstract 27
1. Introduction 28
2. Materials and methods 29 2.1 Lipid extraction of A. ventricosus eggs 29
2.2 Cell proliferation assay 29
2.3 Nitric oxide (NO) assay 29
2.4 Quantitative real-time PCR ( 31
2.5 Protein extraction and western blotting analysis 31
2.6 Fluorescence activated cell sorting (FACS) analysis 32
2.7 Statistical analysis 33
3. Results and discussion 33 3.1 Cytotoxicity of neutral lipids, glycolipids, and phospholipids on LPS-stimulated RAW264.7 macrophages 33
2.2 Inhibitory effects of neutral lipids, glycolipids, and phospholipids on LPS-induced NO production 37
3.3 Inhibitory effects of neutral lipids, glycolipids, and phospholipids on LPS-induced mRNA expression 42
3.4 Inhibitory effects of neutral lipids, glycolipids, and phospholipids on LPS-induced MAPK and NF-κB signaling pathways 46
2.5 Inhibitory effects of neutral lipids, glycolipids, and phospholipids on LPS-induced CD40 expression 50
4. Conclusions 54
4. References 55
Chapter 4. Summary 60
